1. Field of the Invention
The use of fluorescent labels for the determination of analytes in immunoassays has shown substantial promise and numerous immunoassays have been developed which are dependent upon the accurate measurement of the fluorescence from an assay medium. One of the problems with fluorescence is the number of factors which contribute to background values. Included among these facts are light scatter, instrument variation, fluorescent reagent contamination, and endogeneous fluorescence in the sample.
The endogenous fluorescence can be a serious factor in diminishing the quantitative character of the assay. In many situations, the endogenous fluorescence will vary from sample to sample and will be different in the analyte samples from the standards or calibrators which are employed to provide for translating the observed fluorescent signal into the concentration of the analyte. In order to enhance the accuracy of the assay, it is desirable to diminish or completely remove the contribution of the endogenous fluorescent material to the observed signal during the immunoassay.
2. Description of the Prior Art
Illustrative immunoassays employing fluorescers as a label may be found in U.S. Pat. Nos. 3,998,943; 3,939,350; 3,996,345; and patents cited therein. Helman, et al., Clin. Chem. 20 1193, (1974) published the use of basic hydrogen peroxide for decolorizing hemolyzed and jaundiced serum samples. For background material see, Guilbault, "Practical Fluorescence," Marcel Dekker, New York (1973). An article on the fluorescence of bilirubin is Roth, M., Clin. Chim. ACTA, 17:48F (1967).